Overview

  • Product name
    Anti-CD3G antibody [EPR4517]
    See all CD3G primary antibodies
  • Description
    Rabbit monoclonal [EPR4517] to CD3G
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IP, IHC-P, Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide within Human CD3G (C terminal). The exact sequence is proprietary.
    Database link: P09693

  • Positive control
    • WB: Jurkat and mouse thymus lysates. IHC-P: Human tonsil, human colon and mouse spleen tissues. ICC/IF: Jurkat cells. IP: Jurkat cell lysate. FC: Jurkat cells
  • General notes

    Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer
    pH: 7.40
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    EPR4517
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab134096 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/20000 - 1/200000. Predicted molecular weight: 20 kDa.

For unpurified use at 1/10000 - 1/50000.

IP 1/40.

For unpurified use at 1/10 - 1/100.

IHC-P 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

For unpurified use at 1/250 - 1/500.

Flow Cyt 1/70.
ICC/IF 1/1000.

For unpurified use at 1/500 - 1/1000.

Target

  • Function
    The CD3 complex mediates signal transduction.
  • Sequence similarities
    Contains 1 Ig-like (immunoglobulin-like) domain.
    Contains 1 ITAM domain.
  • Domain
    A di-leucine motif and a tyrosine-based motif are individually sufficient to induce both endocytosis and delivery to lysosomes.
  • Cellular localization
    Membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • CD3 gamma antibody
    • CD3-GAMMA antibody
    • CD3g antibody
    • CD3g antigen gamma polypeptide (TiT3 complex) antibody
    • CD3g antigen gamma polypeptide antibody
    • CD3g molecule gamma (CD3 TCR complex) antibody
    • CD3g molecule gamma antibody
    • CD3G_HUMAN antibody
    • FLJ17620 antibody
    • FLJ17664 antibody
    • FLJ79544 antibody
    • FLJ94613 antibody
    • MGC138597 antibody
    • T cell antigen receptor complex gamma subunit of T3 antibody
    • T-cell receptor T3 gamma chain antibody
    • T-cell surface glycoprotein CD3 gamma chain antibody
    • T3G antibody
    see all

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling CD3G with purified ab134096 at 1/1000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • All lanes : Anti-CD3G antibody [EPR4517] (ab134096) at 1/50000 dilution (purified)

    Lane 1 : Jurkat cell lysate
    Lane 2 : Human fetal thymus lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size: 20 kDa
    Observed band size: 18-28 kDa
    why is the actual band size different from the predicted?



    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Immunocytochemistry/Immunofluorescence analysis of Jurkat cells labelling CD3G with purified ab134096 at 1/1000. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. An Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

    Control: primary antibody (1/1000) and secondary antibody, ab150113, an Alexa Fluor® 488-conjugated goat anti-mouse IgG (1/500).

  • Flow Cytometry analysis of Jurkat (human acute T cell leukemia) cells labeling CD3G with purified ab134096 at 1/70 dilution (10ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor®488)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.

  • Anti-CD3G antibody [EPR4517] (ab134096) at 1/50000 dilution (purified) + Mouse thymus tissue lysate at 20 µg

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size: 20 kDa
    Observed band size: 18-28 kDa why is the actual band size different from the predicted?



    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • All lanes : Anti-CD3G antibody [EPR4517] (ab134096) at 1/10000 dilution (unpurified)

    Lane 1 : Jurkat cell lysate
    Lane 2 : Mouse thymus tissue lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

    Predicted band size: 20 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling CD3G with unpurified ab134096.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse spleen tissue labelling CD3G with unpurified ab134096.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon (lymphocytes) tissue labelling CD3G with unpurified ab134096.

  • Immunohistochemistry (Formalin/PFA-fixed parffin-embedded sections) analysis of human skeletal muscle tissue. Unpurified ab32362 shows negative staining.

  • Immunohistochemistry (Formalin/PFA-fixed parffin-embedded sections) analysis of human brain tissue. Unpurified ab32362 shows negative staining.

  • ab134096 (purified) at 1/40 immunoprecipitating CD3G in Jurkat cell lysate (Lane 1). Lane 2 - PBS. For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

References

This product has been referenced in:
  • Ma Y  et al. ACF7 regulates inflammatory colitis and intestinal wound response by orchestrating tight junction dynamics. Nat Commun 8:15375 (2017). Read more (PubMed: 28541346) »
  • Coelho MA  et al. Oncogenic RAS Signaling Promotes Tumor Immunoresistance by Stabilizing PD-L1 mRNA. Immunity 47:1083-1099.e6 (2017). Read more (PubMed: 29246442) »
See all 10 Publications for this product

Customer reviews and Q&As

Application
Flow Cytometry
Fixation
Paraformaldehyde
Sample
Human Cell (Jurkat cells (T lymphocytes))
Specification
Jurkat cells (T lymphocytes)
Permeabilization
Yes - SAPONIN

Ms. Beatriz Garcillan

Verified customer

Submitted Oct 30 2013

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